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Focusing on the research and development and application of isothermal nucleic acid amplification technology

2020年11月 禽流感RT-RAA侧向流试纸条可视化检测方法的建立-河北农大

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Avian influenza (AI) is a major disease that poses a significant threat to the healthy development of China's poultry industry. To address this challenge, this study aimed at developing an efficient and convenient detection method for AI. In this experiment, primers and probes were designed based on the conserved NP gene sequence of avian influenza virus (AIV). Using reverse transcription-recombinase polymerase amplification (RT-RAA) technology, the NP gene was rapidly amplified under isothermal conditions, generating a dual-labeled product. The amplified product was then detected via a lateral flow dipstick (LFD) assay. By optimizing key parameters such as reaction time, temperature, and primer concentration, we successfully established an RT-RAA-LFD detection method capable of identifying avian influenza virus. The results demonstrated that, under 37°C conditions, the target NP gene fragment could be effectively amplified within 24 minutes. The resulting amplicon was easily visualized with the naked eye using the lateral flow dipstick. Importantly, the method exhibited excellent specificity, showing no cross-reactivity with nucleic acids from infectious bronchitis virus (IBV), avian infectious laryngotracheitis virus (AILV), or Newcastle disease virus (NDV). Moreover, the sensitivity of this assay reached 102 copies/μL—100 times higher than conventional PCR—and its clinical detection accuracy was 94.8%. Overall, this method boasts high specificity, exceptional sensitivity, and remarkable convenience and speed, making it a practical tool for field-based detection of avian influenza virus.

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2016年9月 重组酶介导的等温扩增法检测 H7N9禽流感病毒-江苏疾控

多杀性巴氏杆菌荧光RAA快速检测方法的建立-杜秋明

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2016年9月 重组酶介导的等温扩增法检测 H7N9禽流感病毒-江苏疾控

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多杀性巴氏杆菌荧光RAA快速检测方法的建立-杜秋明